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Biomedical subjects

B Herman

Publications and source records attributed to B Herman.

At least 145 records · Page 8Linked to original sources
PubMed1986

Analysis of deoxyribonucleic acid distribution in noncleaving oocytes from patients undergoing in vitro fertilization.

The purpose of this study was to describe the quantity and distribution of deoxyribonucleic acid (DNA) in oocytes that did not fertilize or did fertilize and failed to cleave, from patients who underwent in vitro fertilization. Patients were selected with at least one cleaving egg, so that the sperm population was known to be fertile, and failure of fertilization or cleavage in the remaining oocytes could be attributed to nonspermatozoan factors. The noncleaving oocytes were classified into five categories, the majority of which (71%) lacked a polar body and any morphologically identifiable nucleus or germinal vesicle. Three general defects were found: failure to replicate the DNA properly; failure to package the DNA properly; and failure to organize the nuclear material properly after sperm penetration. It is concluded that either altered stimulation protocols or altered in vitro maturation conditions are needed to increase the average number of normal embryos available for transfer.

Cleavage Stage, Ovum↗
PubMed1985

Binding and internalization of heparin by vascular smooth muscle cells.

Previous work from our laboratory has demonstrated that heparin specifically inhibits the proliferation of vascular smooth muscle cells in vivo and in vitro. In this paper, we examine the binding and mode of internalization of heparin by smooth muscle cells. For these studies, radiolabeled and fluoresceinated (FITC) heparin probes were synthesized that retained their antiproliferative capacity. Binding of 3H-heparin to these cells occurs via specific, high-affinity binding sites (Kd = 10(-9) M, 100,000 binding sites per cell). Approximately 80% of the heparin bound to the cell surface was shed into the culture medium within 2 hr. The heparin that was left on the cell surface was internalized with biphasic kinetics. Approximately 50% of the bound material was internalized within 2 hr. After this initial rapid uptake, the rate slowed substantially, with the remaining heparin requiring 1-2 days to be internalized. Binding and uptake of FITC heparin was monitored using video image intensification fluorescence microscopy. When smooth muscle cells were exposed to FITC heparin at 4 degrees C, a diffuse surface staining pattern was observed. After warming the cells to 37 degrees C, intensely fluorescent vesicles were seen superimposed over the diffuse surface staining within 2 min. After 15 min at 37 degrees C, numerous large punctate vesicles were seen inside the cell. After 2 hr these vesicles had concentrated in the perinuclear region. This pattern of uptake, when considered along with the presence of specific, high-affinity binding sites and the initial rapid uptake of 3H-heparin, suggests that heparin enters smooth muscle cells by both receptor-mediated and other endocytic pathways.

Animals↗
PubMed1985

[Instrumental esophageal perforation--its treatment and results].

The number of oesophagus perforations has increased considerably with the increase in endoscopic, diagnostic and therapeutic procedures. 46 instrumental oesophagus perforations have been observed between 1973 and 1984. The youngest patient was a 2-year-old girl with a benign oesophagus stenosis and the eldest was an 89-year-old man with a neoplasm of the cardia. The average age was 62 years. 26 patients were treated conservatively; 20 patients were treated surgically. In 2 cases the oesophagus was diverted at the cervical level and the cardia detached. 8 months later this was successfully reconstructed. 9 out of 43 patients died. In every case, the cause of death were cardio-pulmonal complications accompanying general sepsis.

Adolescent↗
PubMed1985

Platelet-derived growth factor-induced alterations in vinculin and actin distribution in BALB/c-3T3 cells.

Exposure of BALB/c-3T3 cells (clone A31) to platelet-derived growth factor (PDGF) results in a rapid time- and dose-dependent alteration in the distribution of vinculin and actin. PDGF treatment (6-50 ng/ml) causes vinculin to disappear from adhesion plaques (within 2.5 min after PDGF exposure) and is followed by an accumulation of vinculin in punctate spots in the perinuclear region of the cell. This alteration in vinculin distribution is followed by a disruption of actin-containing stress fibers (within 5 to 10 min after PDGF exposure). Vinculin reappears in adhesion plaques by 60 min after PDGF addition while stress fiber staining is nondetectable at this time. PDGF treatment had no effect on talin, vimentin, or microtubule distribution in BALB/c-3T3 cells; in addition, exposure of cells to 5% platelet-poor plasma (PPP), 0.1% PPP, 30 ng/ml epidermal growth factor (EGF), 30 ng/ml somatomedin C, or 10 microM insulin also had no effect on vinculin or actin distribution. Other competence-inducing factors (fibroblast growth factor, calcium phosphate, and choleragen) and tumor growth factor produced similar alterations in vinculin and actin distribution as did PDGF, though not to the same extent. PDGF treatment of cells for 60 min followed by exposure to EGF (0.1-30 ng/ml for as long as 8 h after PDGF removal), or 5% PPP resulted in the nontransient disappearance of vinculin staining within 10 min after EGF or PPP additions; PDGF followed by 0.1% PPP or 10 microM insulin had no effect. Treatment of cells with low doses of PDGF (3.25 ng/ml), which did not affect vinculin or actin organization in cells, followed by EGF (10 ng/ml), resulted in the disappearance of vinculin staining in adhesion plaques, thus demonstrating the synergistic nature of PDGF and EGF. These data suggest that PDGF-induced competence and stimulation of cell growth in quiescent fibroblasts are associated with specific rapid alterations in the cellular organization of vinculin and actin.

Actins↗
PubMed1985

[Esophagus perforation--treatment and results].

Esophageal perforation is most often caused by instrumental lesion during endoscopy. Mortality is still high; underlying malignant disease, septic complications and cardiopulmonary problems are endangering the patient. In small perforation with poor clinical symptoms, conservative treatment (i.e. antibiotics, gastric suction, parenteral nutrition) should be considered. We could treat in this way more than 50% of our cases. On the other hand, some patients require agressive surgical procedures, e.g. cervical esophagostomy gastrostomy and disconnection of esophageal continuity.

Adolescent↗
PubMed1984

A time-lapse video image intensification analysis of cytoplasmic organelle movements during endosome translocation.

Vital fluorescence staining has been used in conjunction with time-lapse video image intensification microscopy to analyze the distribution and movement of endosomes, lysosomes, and mitochondria in cultured rat ovarian granulosa cells. Exposure of 5-d granulosa cell cultures to pyrene-concanavalin A (P-Con A) or 3,3'-dioctadecylindocarbocyanine-labeled low-density lipoprotein (dil-LDL) at 4 degrees C results in the formation of randomly distributed endosomes 10 min after warming to 37 degrees C that exhibit saltatory motion for 20 min. If granulosa cells are labeled at 4 degrees C with both P-Con A and dil-LDL and warmed to 37 degrees C, both ligands are found within the same endosomes which migrate centripetally to the cell center where label accumulates within phase-dense structures by 60 min. The initial endosome saltations occur over short distances (mean distance = 4.6 micron) with a mean velocity of 0.03 micron/s. Endosome saltations then cease and are followed by a gradual centriptal migration of endosomes to the cell center where they accumulate and fuse with phase-dense structures. The second phase of movement involves a continuous, unidirectional migration of endosomes over distances ranging from 5 to 40 micron at a mean velocity of 0.05 micron/s. Lysosomes were simultaneously visualized as acridine orange-staining, phase-dense structures in control cells and cells exposed to fluorescent ligands. In untreated cells, lysosomes are dispersed throughout the cytoplasm and undergo bidirectional saltations covering a mean distance of 8.7 micron with a mean velocity of 0.3 micron/s. Lysosomes redistribute centripetally to the perinuclear region of the cell by saltatory movement within 20 min of exposure to ligand. Mitochondria were visualized with the fluorescent dye rhodamine 123 in granulosa cells labeled with P-Con A and were found to redistribute to the cell center coincident with endosomes. The microtubule-disrupting agent nocodazole was found to inhibit lysosome saltations and all phases of endosome movement. Taxol, a microtubule-stabilizing agent, partially impaired lysosome movement and led to a redistribution of lysosomes into linear aggregates surrounding the nucleus. Taxol was also found to inhibit endosome movement. The data indicate that (a) endosome movement proceeds initially by saltation and later by a nonsaltatory centripetal migration in association with mitochondria, that (b) lysosomes and endosomes undergo a temporally distinct but spatially similar change in cytoplasmic distribution, and that (c) microtubules are required for the directed translocation of endosomes and lysosomes towards the cell center.

Animals↗
PubMed1984

In vivo and in vitro studies on the role of HMW-MAPs in taxol-induced microtubule bundling.

The involvement of high molecular weight microtubule-associated proteins (HMW-MAPs) in the process of taxol-induced microtubule bundling has been studied using immunofluorescence and electron microscopy. Immunofluorescence microscopy shows that HMW-MAPs are released from microtubules in granulosa cells which have been extracted in a Triton X-100 microtubule-stabilizing buffer (T-MTSB), unless the cells are pretreated with taxol. 1.0 microM taxol treatment for 48 h results in microtubule bundle formation and the retention of HMW-MAPs in these cells upon extraction with T-MTSB. Electron microscopy demonstrates that microtubules in control cytoskeletons are devoid of surface structures whereas the microtubules in taxol-treated cytoskeletons are decorated by globular particles of a mean diameter of 19.5 nm. The assembly of 3 X cycled whole microtubule protein (tubulin plus associated proteins) in vitro in the presence of 1.0 microM taxol, results in the formation of closely packed microtubules decorated with irregularly spaced globular particles, similar in size to those observed in cytoskeletons of taxol-treated granulosa cells. Microtubules assembled in vitro in the absence of taxol display prominent filamentous extensions from the microtubule surface and center-to-center spacings greater than that observed for microtubules assembled in the presence of taxol. Brain microtubule protein was purified into 6 s and HMW-MAP-enriched fractions, and the effects of taxol on the assembly and morphology of these fractions, separately or in combination, were examined. Microtubules assembled from 6 s tubulin alone or 6 s tubulin plus taxol (without HMW-MAPs) were short, free structures whereas those formed in the presence of taxol from 6 s tubulin and a HMW-MAP-enriched fraction were extensively crosslinked into aggregates. These data suggest that taxol induces microtubule bundling by stabilizing the association of HMW-MAPs with the microtubule surface which promotes lateral aggregation.

Alkaloids↗
PubMed1983

Multivariate logistic analysis of risk factors for stroke in Tilburg, The Netherlands.

By means of a case-control study conducted between October 1, 1978, and July 31, 1981, in Tilburg, The Netherlands, various characteristics and events, including personal data, health-related behavior, and medical history, were evaluated as risk factors for stroke. The study subjects included 132 stroke patients and 239 age- and sex-matched control patients interviewed at the two city hospitals. To assess joint effects and possible interactions, and to control for multiple confounding factors, a series of multivariate logistic models for matched data were studied. From this analysis, it appeared that hypertension, acute myocardial infarction, cardiac arrhythmias, transient cerebral ischemic attacks, obesity, physical activity during leisure time, education of head of household, and Rhesus factor were all significant stroke risk factors. These risk determinants demonstrated a multiplicative effect in general; however, the influence of some variables on stroke risk was not constant with age (hypertension, acute myocardial infarction, cardiac arrhythmias, obesity, and Rhesus factor) and sex (hypertension and education of head of household). The relationship of diabetes mellitus to stroke slightly decreased and became nonsignificant after adjustment for factors besides age and sex. Stroke risk was not associated with cigarette and alcohol use, family history of stroke and related disorders, marital status, and ABO blood typing.

Adult↗
PubMed1983

Microtubule regulation of cell surface receptor topography during granulosa cell differentiation.

A possible role for cytoplasmic microtubules in modulating lectin binding site topography has been examined during the hormone-directed differentiation of rat ovarian granulosa cells in vitro. Indirect immunofluorescence staining with anti-tubulin antibodies indicates that undifferentiated cultured granulosa cells contain a network of microtubules which radiate from the cell center to the cell periphery. Cultures induced to differentiate by a three day treatment with 1 microgram/ml prolactin exhibit a marginal distribution of microtubules and a centrally-located primary cilium. Prolactin enhances the incidence of granulosa cells containing a primary cilium from 9% in undifferentiated cultures to 53% in hormone-treated cultures. The pattern of lectin binding site redistribution induced by Concanavalin A (Con A) is also modified by prolactin treatment. In contrast to undifferentiated cells, which randomly endocytose fluorescein Con A, granulosa cells exposed to prolactin respond to fluorescein Con A by forming central surface caps to a greater extent (75%) than undifferentiated controls (25%). Double label fluorescence microscopy and transmission electron microscopy on Con A labeled cells show that caps form at central cell surface sites which contain the primary cilium. Disruption of cytoplasmic microtubules by colchicine, in undifferentiated granulosa cells, results in the formation of cell surface caps upon Con A addition. These data suggest that cytoplasmic microtubules modulate the topography of lectin bindings sites which is subject to hormonal control during the in vitro differentiation of ovarian granulosa cells.

Animals↗
PubMed1983

The effects of taxol on the organization of the cytoskeleton in cultured ovarian granulosa cells.

Exposure of ovarian granulosa cells to taxol, a potent microtubule assembly promoting and stabilization agent, results in a time and dose-dependent alteration in the organization of cytoplasmic filaments (microtubules, microfilaments, intermediate filaments) and organelles. Transmission electron microscopy and fluorescence microscopy are used in conjunction with various antimitotic drugs to evaluate the action of taxol on suspension and monolayer cultures. Taxol treatment (1.0 microM) of freshly isolated suspended cells for 4 h leads to the formation of multiple bundles of microtubules emanating from a single centrosomal organizing center. Within 4 h after addition to monolayer cultures, 1.0 microM taxol induces a lateral aggregation of microtubules which, by 12 to 24 h of treatment, results in the appearance of dense bundles of tightly-packed microtubules located near the centrosome, close to the outer nuclear envelope, and at peripheral cytoplasmic sites. By electron microscopy, both lateral and end-on associations between bundled microtubules and the nuclear envelope are apparent. During the course of taxol-induced bundling, intermediate filaments are altered from a normally dispersed fibrous network into perinuclear aggregates. Microtubule bundling is also associated with a rearrangement of actin filaments from stress fibers into a marginal distribution, and actin appears to be excluded from sites of bundle formation. When granulosa cells are treated with equivalent concentrations of the microtubule-disrupting drugs, colchicine or nocodazole, either before or during taxol treatment, bundle formation is prevented suggesting that an intact microtubule network is required for taxol-induced bundling. Colchicine, but not nocodazole, is able to reverse the effects of taxol on bundle formation. The data suggest that the cellular distribution of cytoplasmic organelles, intermediate filaments, and microfilaments are regulated by the cytoplasmic microtubule complex.

Alkaloids↗
PubMed1982

The intracellular movement of endocytic vesicles in cultured granulosa cells.

Ligand binding to cell surface receptors induces rapid internalization of ligand-receptor complexes by receptor mediated endocytosis. We have examined the intracellular movement of endocytic vesicles, induced by the lectin concanavalin A (Con A), in cultured rat ovarian granulosa cells using fluorescence and electron microscopy. Within 20 minutes of ligand treatment at 37 degrees C, numerous Con A-containing endocytic vesicles form, which migrate to the cell center by 60 minutes. Double label fluorescence microscopy, using fluorescein-Con-A and rhodamine immunofluorescent staining of tubulin or vimentin, indicates that during vesicle migration microtubules and 10-nm filaments are altered in their organization. By 30 minutes, vesicles are associated with microtubule bundles, which subsequently collapse around the nucleus. Similarly, 10-nm filaments accumulate around the nucleus in conjunction with the perinuclear aggregation of endocytic vesicles. Electron microscopy of Con A-horseradish peroxidase-labeled cells demonstrates that endocytic vesicles fuse to form large receptosome-like structures during intracellular migration and these structures are associated with cytoplasmic microtubules and 10-nm filaments. Taxol, a drug that stabilizes microtubules, prevents endocytic vesicle translocation to the Golgi region. Nocodazole, which causes microtubule disassembly, results in the collapse of 10-nm filaments and the central aggregation of endocytic vesicles. The data indicate that the cytoskeleton participates in the directed intracellular movement of endocytic vesicles; the possible subcellular basis for this movement is discussed.

Alkaloids↗